Fluorescence in situ hybridization (FISH) has been employed as a molecular technique to detect DNA sequences on human chromosomes for more than 20 years (Bauman 1985, Acta Histochem Suppl. 31:9-18; Pinkel 1986, PNAS 83(9):2934-8). Over the past two decades, refinement of various aspects of the FISH technique has advanced the field of human cytogenetics and molecular diagnostics, allowing for the identification of chromosomal abnormalities associated with solid tumors and hematopoietic malignancies, and for the diagnosis of infectious diseases. (Heim and Mitelman 1995, Genes Chromosomes Cancer 14(1):56-62; Klinger 1995, Prenat Diagn. 15(1):1-5; Timm, Podniesinski et al. 1995, Cytometry 22(3):250-5; Heselmeyer, Macville et al. 1997, Genes Chromosomes Cancer 19(4):233-40; Sauer, Wiedswang et al. 2003, APMIS 111(3):444-50).
Current FISH procedures are labor intensive and time consuming, requiring multiple manual processing steps and adherence to precise temperature and time requirements. Standard FISH techniques typically require more than a dozen steps to process a slide sample, several of which are performed at different temperatures, necessitating the use of numerous, and often costly, temperature equipment, such as water baths, hot plates, and incubators. These and other limitations of the technique have prevented it from being used more widely in research and clinical laboratories.
Presently, there is a need for more simplified and cost-effective methods of performing FISH that require fewer processing steps, less time and less equipment.